Abstract
The prolamellar body (PLB) proteome of dark-grown wheat leaves was characterized. PLBs are formed not only in etioplasts but also in chloroplasts in young develo** leaves during the night, yet their function is not fully understood. Highly purified PLBs were prepared from 7-day-old dark-grown leaves and identified by their spectral properties as revealed by low-temperature fluorescence spectroscopy. The PLB preparation had no contamination of extra-plastidal proteins, and only two envelope proteins were found. The PLB proteome was analysed by a combination of 1-D SDS-PAGE and nano-LC FTICR MS. The identification of chlorophyll synthase in the PLB fraction is the first time this enzyme protein was found in extracts of dark-grown plants. This finding is in agreement with its previous localization to PLBs using activity studies. NADPH:protochlorophyllide oxidoreductase A (PORA), which catalyses the reduction of protochlorophyllide to chlorophyllide, dominates the proteome of PLBs. Besides the identification of the PORA protein, the PORB protein was identified for the first time in dark-grown wheat. Altogether 64 unique proteins, representing pigment biosynthesis, photosynthetic light reaction, Calvin cycle proteins, chaperones and protein synthesis, were identified. The in number of proteins’ largest group was the one involved in photosynthetic light reactions. This fact strengthens the assumption that the PLB membranes are precursors to the thylakoids and used for the formation of the photosynthetic membranes during greening. The present work is important to enhance our understanding of the significance of PLBs in chloroplast development.
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Abbreviations
- 1-D:
-
One-dimensional
- 2-D:
-
Two-dimensional
- comp-PLB:
-
Comparative-PLB
- EPIM:
-
Etioplast inner membrane
- nano-LC:
-
Nanoliquid chromatography
- FTICR MS:
-
Fourier transform-ion cyclotron resonance mass spectrometry
- Pchlide:
-
Protochlorophyllide
- PLB:
-
Prolamellar body
- POR:
-
NADPH:protochlorophyllide oxidoreductase
- PT:
-
Prothylakoid
- SDS-PAGE:
-
Sodium dodecyl sulphate polyacrylamide gel electrophoresis
References
Armstrong GA, Runge S, Frick G, Sperling U, Apel K (1995) Identification of NADPH:protochlorophyllide oxidoreductases A and B: a branched pathway for light-dependent chlorophyll biosynthesis in Arabidopsis thaliana. Plant Physiol 108:1505–1517
Aronsson H, Jarvis P (2002) A simple method for isolating import-competent Arabidopsis chloroplasts. FEBS Lett 529:215–220
Baginsky S, Kleffmann T, von Zychlinski A, Gruissem W (2005) Analysis of shotgun proteomics and RNA profiling data from Arabidopsis thaliana chloroplasts. J Proteome Res 4:637–640
Bahl J, Francke B, Monéger R (1976) Lipid composition of envelopes, prolamellar bodies and outer plastid membranes in etiolated, green and greening wheat leaves. Planta 129:193–201
Benli M, Schulz R, Apel K (1991) Effect of light on the NADPH-protochlorophyllide oxidoreductase of Arabidopsis thaliana. Plant Mol Biol 16:615–625
Birve SJ, Selstam E, Johansson LB-Å (1996) Secondary structure of NADPH:protochlorophyllide oxidoreductase examined by circular dichroism and prediction methods. Biochem J 317:549–555
Blomqvist LA, Ryberg M, Sundqvist C (2006) Proteomic analysis of the etioplast inner membranes of wheat (Triticum aestivum) by two-dimensional electrophoresis and mass spectrometry. Physiol Plant 128:368–381
Bruce BD (1998) The role of lipids in plastid protein import. Plant Mol Biol 38:223–246
Böddi B, Lindsten A, Ryberg M, Sundqvist C (1989) On the aggregational states of protochlorophyllide and its protein complexes in wheat etioplasts. Physiol Plant 76:135–143
Böddi B, Evertsson I, Ryberg M, Sundqvist C (1996) Protochlorophyllide transformation and chlorophyll accumulation in epicotyls of pea (Pisum sativum). Physiol Plant 96:706–713
Carlsohn E, Nyström J, Karlsson H, Svennerholm A-M, Nilsson CL (2006) Characterization of the outer membrane protein profile from disease-related Helicobacter pylori isolates by subcellular fractionation and nano-LC FT-ICR MS analysis. J Proteome Res 5:3197–3204
Chen S, Harmon AC (2006) Advances in plant proteomics. Proteomics 6:5504–5516
Domanskii V, Rassadina V, Gus-Mayer S, Wanner G, Schoch S, Rüdiger W (2003) Characterization of two phases of chlorophyll formation during greening of etiolated barley leaves. Planta 216:475–483
Donnelly BE, Madden RD, Ayoubi P, Porter DR, Dillwith JW (2005) The wheat (Triticum aestivium L.) leaf proteome. Proteomics 5:1624–1633
Ferro M, Salvi D, Brugière S, Miras S, Kowalski S, Louwagie M, Garin J, Joyard J, Rolland N (2003) Proteomics of the chloroplast envelope membranes from Arabidopsis thaliana. Mol Cell Proteomics 2:325–345
Friso G, Giacomelli L, Ytterberg J, Peltier J-B, Rudella A, Sun Q, van Wijk KJ (2004) In-depth analysis of the thylakoid membrane proteome of Arabidopsis thaliana chloroplasts: new proteins, new functions, and a plastid proteome database. Plant Cell 16:478–499
Froelich JE, Wilkerson CG, Ray WK, McAndrew RS, Osteryoung KW, Gage DA, Phinney BS (2003) Proteomic study of the Arabidopsis thaliana chloroplastic envelope membrane utilizing alternatives to traditional two-dimensional electrophoresis. J Proteome Res 2:413–425
Funk C, Schröder WP, Napiwotzki A, Tjus SE, Renger G, Andersson B (1995) The PSII-S protein of higher plants: a new type of pigment-binding protein. Biochemistry 34:11133–11141
Gaubier P, Wu H-J, Laudié M, Delseny M, Grellet F (1995) A chlorophyll synthetase gene from Arabidopsis thaliana. Mol Gen Genet 249:58–64
Grevby C, Engdahl S, Ryberg M, Sundqvist C (1989) Binding properties of NADPH-protochlorophyllide oxidoreductase as revealed by detergent and ion treatments of isolated and immobilized prolamellar bodies. Physiol Plant 77:493–503
Gunning BES, Steer MW (1996) Plant cell biology structure and function. Jones and Bartlett Publishers International, London
Hashimoto A, Akasaka T, Yamamoto Y (1993) Characteristics of the assembly of the 33 kDa oxygen-evolving complex protein in the etioplasts and the develo** chloroplasts of barley seedlings. Biochim Biophys Acta 1183:397–407
Hinz G, Hillmer S, Bäumer M, Hohl I (1999) Vacuolar storage proteins and the putative vacuolar sorting receptor BP-80 exit the golgi apparatus of develo** pea cotyledons in different transport vesicles. Plant Cell 11:1509–1524
Holtorf H, Reinbothe S, Reinbothe C, Bereza B, Apel K (1995) Two routes of chlorophyllide synthesis that are differentially regulated by light in barley (Hordeum vulgare L.). Proc Natl Acad Sci USA 92:3254–3258
Horton P, Ruban A (2004) Molecular design of the photosystem II light-harvesting antenna: photosynthesis and photoprotection. J Exp Bot 56:365–373
Ikeuchi M, Murakami S (1982) Measurement and identification of NADPH:protochlorophyllide oxidoreductase solubilised with Triton X-100 from etioplast membranes of squash cotyledons. Plant Cell Physiol 23:1089–1099
Joyard J, Block M, Pineau B, Albrieux C, Douce R (1990) Envelope membranes from mature spinach chloroplasts contain a NADPH:protochlorophyllide reductase on the cytosolic side of the outer membrane. J Biol Chem 265:21820–21827
Jürgens G (2004) Membrane trafficking in plants. Annu Rev Cell Dev Biol 20:481–504
Kleffmann T, Russenberger D, von Zychlinski A, Christopher W, Sjolander K, Gruissem W, Baginsky S (2004) The Arabidopsis thaliana chloroplast proteome reveals pathway abundance and novel protein functions. Curr Biol 14:354–362
Kleffmann T, von Zychlinski A, Russenberger D, Hirsch-Hoffmann M, Gehrig P, Gruissem W, Baginsky S (2007) Proteome dynamics during plastid differentiation in rice. Plant Physiol 143:912–923
Kósa A, Márton Z, Böddi B (2005) Fast phototransformation of the 636 nm-emitting protochlorophyllide form in epicotyls of dark-grown pea (Pisum sativum). Physiol Plant 124:132–142
Kroll D, Meierhoff K, Bechtold N, Kinoshita M, Westphal S, Votchknecht UC, Soll J, Westhoff P (2001) VIPP1, a nuclear gene of Arabidopsis thaliana essential for thylakoid membrane formation. Proc Natl Acad Sci USA 98:4238–4242
Kuroda H, Masuda T, Ohta H, Shioi Y, Takamiya K (1996) Effects of light, developmental age and phytohormones on the expression of the gene encoding NADPH-protochlorophyllide oxidoreductase in Cucumis sativus. Plant Physiol Biochem 34:17–22
Lindsten A, Ryberg M, Sundqvist C (1988) The polypeptide composition of highly purified prolamellar bodies and prothylakoids from wheat (Triticum aestivium) as revealed by silver staining. Physiol Plant 72:167–176
Lindsten A, Welch CJ, Schoch S, Ryberg M, Rüdiger W, Sundqvist C (1990) Chlorophyll synthetase is latent in well preserved prolamellar bodies of etiolated wheat. Physiol Plant 80:277–285
Lindsten A, Wiktorsson B, Ryberg M, Sundqvist C (1993) Chlorophyll synthetase activity is relocated from transforming prolamellar bodies to develo** thylakoids during irradiation of dark-grown wheat. Physiol Plant 88:29–36
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the folin phenol reagent. J Biol Chem 193:265–275
Lütz C, Röper U, Beer NS, Griffiths T (1981) Sub-etioplast localization of the enzyme NADPH:protochlorophyllide oxidoreductase. Eur J Biochem 118:347–353
Masuda T, Fusada N, Oosawa N, Takamatsu K, Yamamoto YY, Ohto M, Nakamura K, Goto K, Shibata D, Shirano Y, Hayashi H, Kato T, Tabata S, Shimada H, Ohta H, Takamiya K (2003) Functional analysis of isoforms of NADPH:protochlorophyllide oxidoreductase (POR), PORB and PORC, in Arabidopsis thaliana. Plant Cell Physiol 44:963–974
Morré DJ, Selldén G, Sundqvist C, Sandelius AS (1991) Stromal low temperature compartment derived from the inner membrane of the chloroplast envelope. Plant Physiol 97:1558–1564
Müller B, Eichacker LA (1999) Assembly of the D1 precursor in monomeric photosystem II reaction center precomplexes precedes chlorophyll a-triggered accumulation of reaction center II in barley etioplasts. Plant Cell 11:2365–2377
Olsen JV, Ong SE, Mann M (2004) Trypsin cleaves exclusively C-terminal to arginine and lysine residues. Mol Cell Proteomics 3:608–614
Oster U, Rüdiger W (1997) The G4 gene of Arabidopsis thaliana encodes a chlorophyll synthase of etiolated plants. Bot Acta 110:420–423
Paulsen H (2001) Pigment assembly—transport and ligation In: Aro E-M, Andersson B (eds) Regulation of photosynthesis. Kluwer, Dordrecht, pp 219–233
Peltier JB, Emanuelsson O, Kalume DE, Ytterberg J, Friso G, Rudella A, Liberles DA, Soderberg L, Roepstorff P, von Heijne G, van Wijk KJ (2002) Central functions of the lumenal and peripheral thylakoid proteome of Arabidopsis determined by experimentation and genome-wide prediction. Plant Cell 14:211–236
Peltier JB, Ytterberg AJ, Sun Q, van Wijk KJ (2004) New functions of the thylakoid membrane proteome of Arabidopsis thaliana revealed by a simple, fast, and versatile fractionation strategy. J Biol Chem 279:49367–49383
Pfeffer SR (2007) Unsolved mysteries in membrane traffic. Annu Rev Biochem 76:629–645
Pineau B, Dubertret G, Joyard J, Douce R (1986) Fluorescence properties of the envelope membranes from spinach chloroplasts. J Biol Chem 261:9210–9215
Plücken H, Müller B, Grohmann D, Westhoff P, Eichacker LA (2002) The HCF136 protein is essential for assembly of the photosystem II reaction center in Arabidopsis thaliana. FEBS Lett 532:85–90
Rascio N, Mariani Colombo P, Orsinego M (1980) The ultrastructural development of plastids in leaves of maize plants exposed to continuous illumination. Protoplasma 102:131–139
Rebeiz CC, Rebeiz CA (1986) Chloroplast biogenesis 53: ultrastructural study of chloroplast development during photoperiodic greening. In: Akoyunoglou G, Senger H (eds) Regulation of chloroplast differentiation. AR Liss, New York, pp 389–396
Rossignol M, Peltier J-B, Mock H-P, Matros A, Maldonado AM, Jorrin JV (2006) Plant proteome analysis: a 2004–2006 update. Proteomics 6:5529–5548
Rüdiger W (1993) Esterification of chlorophyllide and its implication for thylakoid development. In: Sundqvist C, Ryberg M (eds) Pigment-protein complexes in plastids: synthesis and assembly. Academic, San Diego, pp 219–240
Rüdiger W, Benz J, Guthoff C (1980) Detection and partial characterization of activity of chlorophyll synthetase in etioplast membranes. Eur J Biochem 109:193–200
Rüdiger W, Böhm S, Helfrich M, Schulz S, Schoch S (2005) Enzymes of the last steps of chlorophyll biosynthesis: modification of the substrate helps to understand the topology of the active center. Biochemistry 44:10864–10872
Ryberg M, Dehesh K (1986) Localization of NADPH-protochlorophyllide oxidoreductase in dark-grown wheat (Triticum aestivum) by immuno-electron microscopy before and after transformation of the prolamellar bodies. Physiol Plant 66:616–624
Ryberg M, Sundqvist C (1982) Characterization of prolamellar bodies and prothylakoids fractionated from wheat etioplasts. Physiol Plant 56:125–132
Ryberg M, Sundqvist C (1988) The regular ultrastructure of isolated prolamellar bodies depends on the presence of membrane-bound NADPH-protochlorophyllide oxidoreductase. Physiol Plant 73:218–226
Ryberg M, Sundqvist C (1991) Structural and functional significance of pigment-protein complexes of chlorophyll precursors. In: Scheer H (ed) Chlorophylls. CRS press, Boca Raton, pp 587–612
Ryrie IJ, Young S, Anderson B (1984) Development of the 33-, 23- and 16-kDa polypeptides of the photosynthetic oxygen-evolving system during greening. FEBS Lett 177:269–273
Schmid HC, Oster U, Kögel J, Lenz S, Rüdiger W (2001) Cloning and characterization of chlorophyll synthase from Avena sativa. Biol Chem 382:903–911
Schoefs B, Franck F (2003) Chlorophyll biosynthesis: mechanisms and evolution. Photochem Photobiol 78:543–557
Schubert M, Petersson UA, Haas BJ, Funk C, Schroder WP, Kieselbach T (2002) Proteome map of the chloroplast lumen of Arabidopsis thaliana. J Biol Chem 277:8354–8365
Selstam E (1998) Development of thylakoid membranes with respect to lipids. In: Siegenthaler P-A, Murata N (eds) Lipids in photosynthesis: structure, function and genetics. Kluwer, Dordrecht, pp 209–224
Shaw P, Henwood J (1985) Immunogold localisation of protochlorophyllide oxidoreductase in barley etioplasts. Eur Cell Biol 39:50–55
Sheramati I, Shahollari B, Landsberger M, Westermann M, Cherepneva G, Kusnetsov V, Oelmüller R (2004) Cytokinin stimulates polyribosome loading of nuclear-encoded mRNAs for the plastid ATP synthase in etioplasts of Lupinus luteus: the complex accumulates in the inner-envelope membrane with the CF1 moiety located towards the stromal space. Plant J 38:578–593
Shevchenko A, Wilm M, Vorm O, Mann M (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem 68:850–858
Solymosi K, Vitányi B, Hideg É, Böddi B (2007) Etiolation symptoms in sunflower (Helianthus annuus) cotyledons partially covered by the pericarp of the achene. Annu Bot 99:857–867
Spano AJ, He Z, Michel H, Hunt DF, Timko MP (1992) Molecular cloning, nuclear gene structure, and developmental expression of NADPH:protochlorophyllide oxidoreductase in pea (Pisum sativum L.) Plant Mol Biol 18:967–972
Sundqvist C, Dahlin C (1997) With chlorophyll pigments from prolamellar bodies to light-harvesting complexes. Physiol Plant 100:748–759
Timko MP (1993) Regulation of gene expression involved in chlorophyll and heme formation in higher plants. In: Lodha ML, Mehta SL, Ramagopal S, Srivastava GP (eds) Advances in plant biotechnology and biochemistry. Indian Soc Agril Biochemists, Kanpur, pp 9–19
Virgin HI (1993) Effectiveness of light of different wavelengths to induce chlorophyll biosynthesis in rapidly and slowly greening tissues. Physiol Plant 89:761–766
von Zychlinski A, Kleffman T, Krishnamurthy N, Sjölander K, Baginsky S, Gruissem W (2005) Proteome analysis of the rice etioplast: metabolic and regulatory networks and novel protein functions. Mol Cell Proteomics 4:1072–1084
Wellburn AR (1982) Bioenergetic and ultrastructural changes associated with chloroplast development. Int Rev Cyt 80:133–191
Wellburn AR, Quail PH, Gunning BES (1977) Examination of ribosome-like particles in isolated prolamellar bodies. Planta 134:45–52
Wellburn AR, Gounaris I, Owen JH, Laybourn-Parry EM, Wellburn FAM (1986) Development of bioenergetic function in light-grown seedlings. In: Akoyunoglou G, Senger H (eds) Regulation of chloroplast differentiation. A.R.Liss, New York, pp 371–381
Widell-Wigge A, Selstam E (1990) Effects of salt wash on the structure of the prolamellar body membrane and the membrane binding of NADPH-protochlorophyllide oxidoreductase. Physiol Plant 78:315–323
Ytterberg J, Peltier J-B, van Wijk KJ (2006) Protein profiling of plastoglobules in chloroplasts and chromoplasts. A surprising site for differential accumulation of metabolic enzymes. Plant Physiol 140:984–997
Zimmer JSD, Monroe ME, Qian W-J, Smith RD (2006) Advances in proteomics data analysis and display using an accurate mass and time tag approach. Mass Spectrom Rev 25:450–482
Acknowledgements
We thank Elisabet Carlsohn and Sjoerd van der Post at the Proteomics Core Facility, Göteborg University, for performing the MS analyses. This work was supported in part by the Royal Society of Arts and Sciences in Göteborg and the Swedish Research Council.
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Blomqvist, L.A., Ryberg, M. & Sundqvist, C. Proteomic analysis of highly purified prolamellar bodies reveals their significance in chloroplast development. Photosynth Res 96, 37–50 (2008). https://doi.org/10.1007/s11120-007-9281-y
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DOI: https://doi.org/10.1007/s11120-007-9281-y